Western Blot: CD36/SR-B3 Antibody - Total protein from Human Skin and Adipose tissue, Mouse Adipose and Rat Adipose tissue was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-CD36 in 5% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Anwendungsinformationen

Western Blot 1:500-1:2000, Immunohistochemistry 1:200-1:400, Immunocytochemistry/Immunofluorescence 1:50-1:200, Immunohistochemistry-Paraffin 1:200-1:400, Immunohistochemistry-FrozenThis CD36 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry paraffin embedded sections, and Western blot analysis where a band is seen ~75-80 kDa. The theoretical molecular weight of CD36 is ~53 kDa. The difference in theoretical MW and actual MW as seen in Western blot is most likely due to the heavy glycosylation and palmitoylation of this protein. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID 24531551) The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Kommentare

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protokoll

Protocol specific for CD36 Antibody Western Blot Protocol1. Perform SDS-PAGE (4-12 %) on samples to be analyzed, loading 50ug of total protein per lane.. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.. Stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.. Rinse the blot in TBST for approximately 5 minutes.. Block the membrane using 5 % non-fat dry milk, 1 % BSA in TBST for 1 hour.. Dilute the rabbit anti-CD36 primary antibody in blocking buffer and incubate overnight at 4 degrees Celcius.. Wash the membrane in TBST 5 times for 5 minutes each.. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.. Wash the blot in TBST 5 times for 5 minutes each.. Apply the detection reagent of choice in accordance with the manufacturers instructions.IHC-FFPE sectionsI. Deparaffinization:A. Treat slides with Xylene: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.B. Treat slides with 100 % Reagent Alcohol: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.II. Quench Endogenous Peroxidase:A. Place slides in peroxidase quenching solution: 15-30 minutes.To Prepare 200 mL of Quenching Solution:Add 3 mL of 30 % Hydrogen Peroxide to 200 mL of Methanol.Use within 4 hours of preparationB. Place slides in distilled water: 2 changes for 2 minutes each.III. Retrieve Epitopes:A. Preheat Citrate

Western Blot: CD36/SR-B3 Antibody - Total protein from Human Skin and Adipose tissue, Mouse Adipose and Rat Adipose tissue was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-CD36 in 5% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.